Comparative neuroimaging of sex differences in human and mouse brain anatomy

  1. Elisa Guma  Is a corresponding author
  2. Antoine Beauchamp
  3. Siyuan Liu
  4. Elizabeth Levitis
  5. Jacob Ellegood
  6. Linh Pham
  7. Rogier B Mars
  8. Armin Raznahan  Is a corresponding author
  9. Jason P Lerch  Is a corresponding author
  1. Section on Developmental Neurogenomics, Human Genetics Branch, National Institute of Mental Health, United States
  2. Mouse Imaging Centre, Canada
  3. The Hospital for Sick Children, Canada
  4. Department of Medical Biophysics, University of Toronto, Canada
  5. Wellcome Centre for Integrative Neuroimaging, Nuffield Department of Clinical 15 Neurosciences, University of Oxford, United Kingdom
  6. Donders Institute for Brain, Cognition and Behaviour, Radboud University Nijmegen, Netherlands
9 figures, 5 tables and 3 additional files

Figures

Figure 1 with 1 supplement
Effects of sex on total tissue volume (TTV) in humans and mice.

Distributions of TTV are shown for the effects of sex in humans (A) and mice (B). Data are represented using individual points, boxplot, and half-violin plot. Linear model used to test for sex differences in each species (correcting for age in both species, Euler number in humans, and background strain in mice) ***p<0.0001 . M=male, F=female.

Figure 1—figure supplement 1
Sex differences in total gray and white matter volume in humans and mice.

Sex differences in total gray matter and total white matter volumes are shown for the humans (A: gray; C: white) and mice (B: gray; D: white). Data are represented using individual points, boxplot, and half-violin plot (raincloud plot). Boxplot midline represents the median, the box represents the first and third quaritles, and the vertical line represent the range of the data. Linear model used to test for sex differences in each species (correcting for age in both species, Euler number in humans, and background strain in mice); ***p<0.001; *p< 0.05; n for humans = 516F/454M, n for mice = 213F/216M.

Figure 2 with 1 supplement
Effect of sex on regional brain volume in humans and mice.

(A, B) Distribution of sex-specific standardized effect sizes across anatomical regions for humans (A) and mice (B). (C, D) Unthresholded (left) and significant (q<0.05; right) standardized effect sizes for the effect of sex displayed on the human (C) and mouse (D) brains. Regions in yellow-red are larger in males and regions in blue are larger in females; n for humans = 516F/454M, n for mice = 213F/216M. Linear model used to test for sex differences in each species across all regions (correcting for age and TTV in both species, Euler number in humans, and background strain in mice). FDR correction used to identify regions with q<0.05.

Figure 2—source data 1

Summary of volumetric sex differences across all regions of the human brain.

The table includes the effect size (positive for male-biased and negative for female-biased), the t-value, p-value, and q-value (from false discovery rate, FDR correction).

https://cdn.elifesciences.org/articles/92200/elife-92200-fig2-data1-v1.xlsx
Figure 2—source data 2

Summary of volumetric sex differences across all regions of the mouse brain.

The table includes the effect size (positive for male-biased and negative for female-biased), the t-value, p-value, and q-value (from false discovery rate, FDR correction).

https://cdn.elifesciences.org/articles/92200/elife-92200-fig2-data2-v1.xlsx
Figure 2—figure supplement 1
Effect of sex on regional brain volume in humans and mice without total tissue volume (TTV) correction.

Distribution of sex-biased standardized effect sizes for humans (A) and mice (B). Unthresholded (left) and significant (q<0.05; right) standardized effect sizes for the effect of sex displayed on the human (C) and mouse (D) brains. Regions in yellow-red are larger in males and regions in blue are larger in females; n for humans = 516F/454M, n for mice = 213F/216M. Linear model used to test for sex differences in each species across all regions (correcting for age in both species, Euler number in humans, and background strain in mice). FDR correction used to identify regions with q<0.05.

Figure 3 with 1 supplement
Sex differences in the variability of regional brain volumes (accounting for TTV differences) in humans and mice.

Distribution of z-scored total brain volume measures across all humans (A) and mouse subjects (B). Uncorrected (P<0.05; left) and significant (q<0.05; right) sex differences in variability (based on Levene’s test) shown on the human brain. Uncorrected (P<0.05) sex differences in variability in the mouse brain (purple = more variable in males; green = more variable in females). Note: all regional human volumes were residualized for TTV, age, and Euler, while regional mouse volumes were residualized for TTV, age, and background strain; n for humans = 516F/454M, n for mice = 213F/216M. Levene’s test for equality of variances used to test for sex differences in variance (corrected for age and TTV in both species, Euler number in humans, and background strain in mice). P-values were corrected with FDR to derive q-values.

Figure 3—figure supplement 1
Sex differences in the variability of regional brain volumes (not accounting for TTV differences) in humans and mice.

Uncorrected (P<0.05; left) and significant (q<0.05; right) sex differences in variability (based on Levene’s test) shown on the human brain (A) Uncorrected (P<0.05) sex differences in variability in the mouse brain (purple = more variable in males; green = more variable in females) (B). Note: all regional human volumes were residualized for age and Euler, while regional mouse volumes were residualized for age and background strain; n for humans = 516F/454M, n for mice = 213F/216M. Levene’s test for equality of variances used to test for sex differences in variance (corrected for age in both species, Euler number in humans, and background strain in mice). P-values were corrected with FDR to derive q-values.

Figure 4 with 1 supplement
Correlation of sex effects on regional volume in homologous regions of the human and murine brain.

(A) Standardized effect size correlation for the effect of sex in humans (x-axis) and mice (y-axis) (robust correlation coefficient, r=0.30). (B) Correlation of standardized effect sizes for the effect of sex across species for cortical regions (green, r=0.31), and non-cortical regions (purple, r=0.16); n for humans = 516F/454M, n for mice = 213F/216M. Robust correlation used to assess correlation between human and mouse sex effect sizes (corrected for TTV and age in both species, Euler number in humans and background strain in mice) yeild r and p values.

Figure 4—figure supplement 1
Correlation of effects of sex in human and mouse homologous brain regions without total tissue volume (TTV) correction.

A. Standardized effect size correlation for the effect of sex in humans (x-axis) and mice (y-axis) (r=0.09). (B). Correlation of standardized effect sizes for the effect of sex across species for cortical regions (green, r=−0.09), and non-cortical regions (purple, r=0.20); n for humans = 516F/454M, n for mice = 213F/216M. Robust correlation used to assess relationship between human and mouse sex effect sizes (corrected for age in both species, Euler number in humans and background strain in mice) yeild r and p values.

Homologous brain regions show either congruent or divergent sex bias across species.

There are no regions that are larger in human females and mouse males (top left quadrant, light green). Several regions show male-bias (i.e., larger volume in males; top right quadrant in yellow) and female-bias (i.e., larger volume in males; females; bottom left quadrant in blue) across species. A subset of regions shows male-bias in humans but female-bias in mice (bottom right quadrant in green). * denotes significant sex effect in both species; n for humans = 516F/454M, n for mice = 213F/216M. Ordering of quadrants matches the quadrants of the scatter plots in Figure 3 and Figure 5.

Figure 6 with 2 supplements
Inter-species anatomical sex congruence and gene expression shows modest correlation across homologous brain regions.

Correlation between similarity in volumetric sex differences and similarity in transcriptional profile using all homologous genes across all homologous regions is modest (A).There was a stronger correlation across cortical (green) compared to non-cortical (purple) regions (B). Robust correlation used to assess correlation between anatomical sex effect similarity and transcriptional similarity yeild r and p values.

Figure 6—figure supplement 1
Similarity matrix for cross-species homologous gene expression.

The matrix displays the correlation between 56 homologous human (x-axis) and mouse (y-axis) brain regions based on the expression of 2835 homologous genes. Hierarchical clustering was used to order the rows and columns using the R function pheatmap; n for humans = 516F/454M, n for mice = 213F/216M. Pearson correlation was used to generate the similarity matrix between all pairs of human and mouse regions; colors denote strenght of pearson correlation coefficient (warm for positive r value, cool for negative r value).

Figure 6—figure supplement 2
Recomputing inter-species anatomical sex congruence and gene expression exlcuding BNST shows similar correlation to analysis including all homologous brain regions.

Correlation between similarity in volumetric sex differences and similarity in transcriptional profile using all homologous genes across but exlcuding the BNST from homologous regions list (A). There was a stronger correlation across cortical (green) compared to non-cortical (purple) regions (B). Robust correlation used to assess correlation between anatomical sex effect similarity and transcriptional similarity yeild r and p values.

Appendix 1—figure 1
Effect of sex on regional brain volume in humans when accounting for relatedness.

Distribution of sex-biased standardized effect sizes for humans when one twin-pair is excluded (A) or when relatedness is accounted for using linear mixed-effects modeling (B). Unthresholded (left) and significant (q<0.05; right) standardized effect sizes for the effect of sex displayed on the human brains. Regions in yellow-red are male-biased and regions in blue are female-biased. (C) Pearson correlation of standardized effect size for sex generated from a linear model with no twin pairs with effect size for sex generated from linear model with twin pairs included (r=0.995). (C) Pearson correlation of standardized effect size for sex generated from a linear model including twin pairs with effect size for sex generated from a the linear mixed-effects model which included twin pairs, but accounting for relatedness by using a family ID as a random effect. Human sample size excluding twin pairs: n=412F/403M; sample size including twin pairs: n=516F/454M.

Appendix 3—figure 1
Robust correlation of anatomical similarity of sex effects and transcriptional similarity of homologous brain regions across species.

Robust correlation of anatomical similarity and transcriptional similarity using only X-chromosome homologous genes (n=91; A, B), using only sex hormone genes androgen, estrogen, progesterone genes (n=30; C, D), just androgen genes (E, F), or just estrogen and progesterone genes (G, H) across all homologous regions or split into cortical and non-cortical.

Appendix 3—figure 2
Generating a null distribution of correlation coefficients for the anatomical vs. transcriptional similarity.

Correlations are significant relative to null distribution (shown in green) generated by randomly sampling subsets of 2835 homologous genes corresponding to the biologically informed subsets, recomputing the transcriptional similarity, and correlating that with the anatomical similarity 10,000 times. We have subsets of 91 genes in (A) corresponding to X-linked genes, 34 genes in (B) corresponding to sex hormone genes, 11 genes in (C) corresponding to androgen games, and 23 genes in (D) corresponding to estrogen and progesterone genes. For each, the observed correlation was compared to the null correlation to generate a p-value, displayed on the graph.

Tables

Table 1
Species-specific effect sizes for volumetric sex differences for 60 homologous brain regions.

Effect sizes are color-coded (blue: larger in males/yellow: larger in females) and asterisk/bold text denotes statistical significance. All results are from analyses covarying for total tissue volume (TTV).

LabelGlasser/Freesurfer# namesMouse atlasHemisphereHuman effect size (β)Mouse effect size (β)
Agranular insulaAVI, AAIC, MIAgranular insular areaL0.200 *–0.558 *
R0.164 *–0.522 *
AmygdalaAmygdala#Cortical subplateL0.305 *0.163 *
R0.201 *0.151 *
Anterior cingulate areaA24pr, a24, p24pr, p24, 24dd, 24dv, p32pr, d32, a32pr, p32, s32Anterior cingulate areaL–0.102 *–0.198
R–0.113 *–0.267 *
Bed nucleus of stria terminalisBed nucleus of stria terminalisBed nucleus of stria terminalisL0.466 *0.918 *
R0.360 *0.971 *
CaudoputamenCaudate#, Putamen#CaudoputamenL0.093–0.224 *
R0.059–0.188 *
Cerebellar cortexCerebellar cortex#Cerebellar cortexL0.430 *–0.268 *
R0.478 *–0.250 *
Dentate gyrus, molecular layerDentate gyrus, molecular layerDentate gyrus, molecular layerL–0.0290.247 *
R0.0410.232 *
CA1CA1CA1L0.151 *0.385 *
R0.1090.377 *
CA3CA3CA3L0.0040.307 *
R0.0040.411 *
Entorhinal cortexECEntorhinal areaL0.470 *–0.090
R0.567 *–0.138
Globus pallidusGlobus Pallidus#PallidumL0.154 *0.112 *
R0.138 *0.180 *
HippocampusHippocampus#Hippocampal regionL0.120 *0.353 *
R0.129 *0.379 *
HypothalamusHypothalamusHypothalamusL0.631 *0.185 *
R0.617 *0.109 *
Medial amygdalar nucleusMedial amygdalar nucleusMedial amygdalar nucleusL0.253 *0.906 *
R0.183 *1.034 *
Medial preoptic areaMedial preoptic areaMedial preoptic areaL0.636 *0.435 *
R0.680 *0.367 *
Nucleus accumbensNucleus accumbens#Striatum ventral regionL–0.311 *–0.005
R–0.249 *0.032
Perirhinal areaPeEc, TF, PHA2, PHA3Perirhinal areaL0.033–0.120
R0.086–0.108
Piriform cortexPirPiriform cortexL0.460–0.131
R0.756 *–0.151
Posterior parietal association areas5 m, 5 mv, 5 LPosterior parietal association areasL–0.254 *0.016
R–0.263 *0.039
Primary auditory areaA1Primary auditory areaL–0.163–0.256 *
R–0.182 *–0.209 *
Primary motor area4Primary motor areaL–0.124–0.329 *
R–0.081–0.357 *
Primary somatosensory area1, 2, 3 a, 3bPrimary somatosensory areaL–0.237 *–0.419 *
R–0.219 *–0.241 *
Primary visual areaV1Primary visual areaL0.175 *0.029
R0.199 *–0.102
Retrosplenial areaRSCRetrosplenial areaL0.198 *0.004
R0.182 *0.035
SubiculumPreSSubiculumL0.182 *0.317 *
R–0.0310.338 *
Temporal association areasFFC, PIT, TE1a, TE1p, TE2a, TF, STV, STSvp, STSvaTemporal association areasL0.0570.085
R0.042–0.004
ThalamusThalamus#ThalamusL–0.028–0.098
R–0.060–0.139 *
Ventral orbital area10 r, 10 vVentral orbital areaL0.083–0.209 *
R–0.046–0.171
Brain stem (midline)Brainstem#Midbrain, HindbrainM0.349 *0.200 *
Medulla (midline)MedullaMedullaM0.385 *0.204 *
Midbrain (midline)MidbrainMidbrainM0.423 *0.191 *
Pons (midline)PonsPonsM0.279 *0.065
Table 2
Demographics for human sample.
FemalesMalesStatistics
Sample size516454
AgeMean29.4127.9F(1,1082)=57.13, p=8.67e-14 ***
SD3.683.61
Range22–3622–37
Education (in years)Mean14.9614.84F(1,1082)=1.465, p=0.226
SD1.831.77
Range11–1711–17
Euler numberMean–52.47–58.26F(1,1082)=25.72, p=4.65e-07 ***
SD17.6619.25
Range–126 to –16–136 to –16
ZygosityMonozygotic10249X2=26.281, p=8.33e-06
Dizygotic163131
Not Twin251274
  1. *p<0.01 **p<0.001 for ANOVA test of significant difference between groups (males vs. females). SD=standard deviation.

Table 3
Demographics for mouse sample.

For details regarding the origin of each mouse cohort refer to Appendix 5—table 1.

FemaleMaleStatistics
Sample size213216
Age
Mean62.062.8F(1,70)=0.78, p=0.38
SD7.58.6
Range56–9056–90
Background Strain
C57BL-6J134141
C57BL-6N7975
Mouse Cohort for C57BL6J
A1012X2=5.46, p=0.91
B1515
C2729
D137
E811
F910
G79
H1010
I79
J106
K98
L915
Mouse Cohort for C57BL6N
M1319 X2=5.745, p=0.332
N108
O2513
P99
Q913
R1312
Appendix 2—table 1
Mapping of homologous human-mouse brain regions (not TTV-corrected standardized effect sizes).

Blue shade highlights female-biased regions, while yellow shade highlights male-biased regions. In humans, the subset of homologous regions was all male-biased due to the larger overall brain size in males. In mice, we observed the same patterns of sex-bias as we did in the analyses which contrived for total tissue volume (TTV) except for the right nucleus accumbens showing no sex bias and a male-bias in the pons (both female-biased in the TTV controlled analysis).

LabelGlasser/Freesurfer# namesMouse atlasHemisphereHuman effect sizeMouse effect size
Agranular insulaAVI, AAIC, MIAgranular insular areaL1.025 *–0.484 *
R0.985 *–0.453 *
AmygdalaAmygdala#Cortical subplateL1.194 *0.265 *
R1.135 *0.240 *
Anterior cingulate areaA24pr, a24, p24pr, p24, 24dd, 24dv, p32pr, d32, a32pr, p32, s32Anterior cingulate areaL0.946 *–0.107
R0.958 *–0.173
Bed nucleus of stria terminalisBed nucleus of stria terminalisBed nucleus of stria terminalisL1.160 *0.937 *
R1.131 *0.959 *
CaudoputamenCaudate#, Putamen#CaudoputamenL0.986 *–0.158
R0.993 *–0.132
Cerebellar cortexCerebellar cortex#Cerebellar cortexL1.123 *–0.220 *
R1.192 *–0.218
Dentate gyrus, molecular layerDentate gyrus, molecular layerDentate gyrus, molecular layerL0.885 *0.292 *
R0.935 *0.275 *
CA1CA1CA1L0.936 *0.427 *
R0.925 *0.417 *
CA3CA3CA3L0.593 *0.382 *
R0.663 *0.460 *
Entorhinal cortexECEntorhinal areaL0.925 *0.038
R0.984 *–0.047
Globus pallidusGlobus Pallidus#PallidumL1.040 *0.167 *
R1.049 *0.229 *
HippocampusHippocampus#Hippocampal regionL1.045 *0.414 *
R1.033 *0.426 *
HypothalamusHypothalamusHypothalamusL1.374 *0.245 *
R1.354 *0.170
Medial amygdalar nucleusMedial amygdalar nucleusMedial amygdalar nucleusL0.547 *0.926 *
R0.654 *1.057 *
Medial preoptic areaMedial preoptic areaMedial preoptic areaL1.205 *0.472 *
R1.275 *0.388 *
Nucleus accumbensNucleus accumbens#Striatum ventral regionL0.596 *0.079
R0.709 *0.106
Perirhinal areaPeEc, TF, PHA2, PHA3Perirhinal areaL0.839 *–0.014
R0.821 *–0.029
Piriform cortexPirPiriform cortexL0.988 *–0.020
R1.076 *–0.039
Posterior parietal association areas5 m, 5 mv, 5 LPosterior parietal association areasL0.519 *0.028
R0.554 *0.066
Primary auditory areaA1Primary auditory areaL0.352 *–0.192
R0.357 *–0.183
Primary motor area4Primary motor areaL0.785 *–0.237 *
R0.795 *–0.269 *
Primary somatosensory area1, 2, 3 a, 3bPrimary somatosensory areaL0.743 *–0.332 *
R0.671 *–0.162
Primary visual areaV1Primary visual areaL0.729 *0.098
R0.703 *–0.085
Retrosplenial areaRSCRetrosplenial areaL0.838 *0.010
R0.770 *0.062
SubiculumPreSSubiculumL0.643 *0.370 *
R0.433 *0.382 *
Temporal association areasFFC, PIT, TE1a, TE1p, TE2a, TF, STV, STSvp, STSvaTemporal association areasL1.084 *0.159
R1.060 *0.024
ThalamusThalamus#ThalamusL1.019 *–0.040
R0.994 *–0.087
Ventral orbital area10 r, 10 vVentral orbital areaL0.666 *–0.121
R0.605 *–0.093
Brain stem (midline)Brainstem#Midbrain, HindbrainM1.209 *0.226 *
Medulla (midline)MedullaMedullaM1.111 *0.255 *
Midbrain (midline)MidbrainMidbrainM1.280 *0.194 *
Pons (midline)PonsPonsM1.101 *0.101
Appendix 5—table 1
Information about the origin laboratory of wild-type (WT) mice was included in the study.
Background strainStudy cohort keyOrigin laboratory
C57BL6JAUniversity of Michigan; Dr. Diane Robinson
BKAIST; Dr. Eunjoon Kim
CUniversity of Western Ontario; Dr. Nathalie Berube
DUT Southwestern; Dr. Genevieve Konopka
EDuke University; Dr. Christelle Golzio
FDuke University; Dr. Christelle Golzio
GLost Angeles Children’s Hospital; Dr. Pat Levitt
HColumbia University; Dr. Jeremy Veenstra-VenderWeele
IScripps Research Institute; Dr. Gavin Rumbaugh
JMcMaster University; Dr. Karun Singh
KMcMaster University; Dr. Jane Foster
LUniversity of Toronto Center for Phenogenomics
C57BL6NMThe Hospital for Sick Children; Dr. Lauryl Nutter
NUC Davis - MIND Institute; Dr. Alex Nord
OThe Hospital for Sick Children; Dr. Lauryl Nutter
PUCSD; Dr. Lilia Iakoucheva
QUT Southwestern; Dr. Graig Powell
RUC Davis; Dr. Alexander Nord

Additional files

MDAR checklist
https://cdn.elifesciences.org/articles/92200/elife-92200-mdarchecklist1-v1.docx
Source data 1

Sex hormone gene and sex chromosome lists for both species.

Gene list based on gene ontology search terms (from Bader Lab) used to identify genes associated with sex hormones including androgen, estrogen, and progesterone, as well as X-chromosome genes.

https://cdn.elifesciences.org/articles/92200/elife-92200-data1-v1.xlsx
Source data 2

Homologous sex hormone and sex chromosome genes.

Genes identified in Source data 1 were filtered to only include homologous genes to allow for cross-species comparison.

https://cdn.elifesciences.org/articles/92200/elife-92200-data2-v1.xlsx

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  1. Elisa Guma
  2. Antoine Beauchamp
  3. Siyuan Liu
  4. Elizabeth Levitis
  5. Jacob Ellegood
  6. Linh Pham
  7. Rogier B Mars
  8. Armin Raznahan
  9. Jason P Lerch
(2024)
Comparative neuroimaging of sex differences in human and mouse brain anatomy
eLife 13:RP92200.
https://doi.org/10.7554/eLife.92200.2